A new alignment calibration method of attitude sensors for the precisionattitude determination of a spacecraft based on the extended Kalman filter is proposed.The proposed method is divided into two steps connected in series: the gyro and thestar tracker calibration. For gyro calibration, alignment errors and scale factor errorsare estimated during the calibration maneuver under the assumption of a perfect startracker. Estimation of the alignment errors of the star trackers and compensation ofthe gyro calibration errors are then performed using the measurements includingpayload information. Performance of the proposed method are demonstrated bynumerical simulations.